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                          Immunohistochemical Detection Kit for P185erbB-2/HER-2 Oncoprotein

 

SPECIFICATION AND INSTRUCTION

[Product Name]

Immunohistochemical Detection Kit for P185erbB-2/HER-2 Oncoprotein

[Package]

20 samples per kit

[Intended Use]

For detecting tumor protein p185erbB-2/HER-2 provide the basis of diagnosis in Breast cancer and ovarian cancer .

[Test Principle]

erbB-2/HER-2 is a oncogene, the protein of 185kD(p185),the positive expression of p185  becomed  a significant tumor marker of international generally accepted. This kit uses self-developed Anti-p185erbB-2/HER-2 Monoclonal antibody, and perform Biotin-Avidin IHC , cancer membrane Stained yellow means positive .

[Units of Kit]

(1)  Solution A        5.0ml      3% H2O2

(2)  Solution B        1.2ml      Blocking Solution

(3)  Solution C        1.2ml      Anti-p185erbB-2/HER-2Monoclonal antibody

(4)  Solution D        1.2ml      Biotinylated secondary  antibody                            

(5)  Solution E        1.2ml      Avidin-labeled HRP                        

(6)  DAB             prepared before use

(7)  positive tissue slice  (Purchased youself) : tissue slice of p185erbB-2/HER-2 positive                              

(8)  Instruction.

[Storage]

Stored at 2 ~ 8℃ for 10 months

[Equipment]

Common light microscope

[Sample]

Tissue slice

[Test method]

(1) Paraffin Sections: Trim paraffin blocks as necessary and cut at 4 um (5 um is commonly used).And the frozen sections,should have been used fixation for frozen tissue sections is to immerse the slides in pre-cooled acetone 4℃ for 10 min;

(2) Dry sections, melting Paraffin:Dry sections in incubator for at least 15 min. at -90℃;

(3)Deparaffinization/Rehydration:Hydrate with Xylene thanol and rinse in distilled water,then can perform Immunohistochemistry;

(4) Rinse in PBSH7.4,0.01M Phosphate buffer or 3x5min;

(5) Allow the slide to drain, Shake off excess Liquid with a brisk motion and carefully wipe each slide around the Sections, Dropping A solution cover the tissue Sections ,incubate

10 min at Room temperature. Shake off excess Liquid, Rinse in PBS for 3x5min;

(6) Allow the slide to drain, Shake off excess Liquid with a brisk motion and carefully wipe each slide as before ,apply B solution cover the tissue Sections ,incubate 10 min at Room temperature;

(7) Allow the slide to drain, Shake off excess Liquid with a brisk motion and carefully wipe each slide as before, apply C solution cover the tissue Sections, apply B solution cover the tissue Section. ,incubate for 1 hour at 37℃ or overnight at 4℃chamber. Rinse in PBS for 3x5min; Allow the slide to drain, Shake off excess Liquid with a brisk motion and carefully wipe each slide as before, apply B solution cover the tissue Section, incubate 10 min at Room temperature, Rinse in PBS for 3x5min;

(8)  Allow the slide to drain, Shake off excess Liquid with a brisk motion and carefully wipe each slide as before ,apply E solution cover the tissue Sections ,incubate 10 min at Room temperature, Rinse in PBS for 3x5min;.

(9)  Prepared DAB solution when it been used, dissolve 5mgDAB in 10ml of PBS,and apply A solution 100μl, (uint desired color reaction is observed when monitored with  the  Microscope);

(10) Rinse in distilled water ,or can do karyotin again when necessary; Dehydrate through Gradient ethanol, Clear in xylene, Coverslip with Gum.

[Reference Result]

Reference value: compared to negative tissue slice ,the positive tissue slice should have 5% cancer membrane Stained yellow, immune reactions were continuous and linear around cell.

[Interpretation of the Results Identified]

Negative: the tissue Sections without stained, <5% cancer membrane Stained yellow, Staining intensity is Low.  

Low positive (±-+) :> 5% cancer membrane Stained yellow, immune reactions were not continuous and linear around cell . Staining intensity is Low,part of cell Stained.

Moderate positive(++): > 5% cancer membrane Stained yellow, immune reactions were continuous and linear around cell        

High positive(+++): > 5% cancer membrane Stained dense yellow, immune reactions were continuous and linear around cell.++-+++ means over-expression

Precaution: The stained Cytoplasm can not be judged for Specific staining .

[Limitations of Test Methods]

Only for tissue slice

[Product Performance]

Comparison with similar products, Correlation coefficient0.654<0.0001

[Precaution]

(1) By use this kit,the  Sections without prior enzyme digestion or antigen rerival.

(2) All solutions applied should  cover the tissue Sections

(3) Room temperature mean 20 ?, Bath at 37 ? better  

(4) pH7.4, 0.01MPBS and DAB need Prepared before use by youself

[Manufactured by]

Anhui Anke Biotechnology (Group) Co., Ltd.

[Address]

Ankebio Buildings, 669, Changjiang Road, West ,Hefei, 230088, P.R. China.

Tel: +86-551-65319890, 65318811

Fax:+86-551-65319895

Website: www.xmobk.live

E-mail: [email protected]

[Medical Device Manufacturing Enterprise License Number] WSYJX 20070120

[The registration Number of Medical Devices] GSYJX(Z)Z2009N3400543

[Standard Number]  YBS00392004

[Instruction Approval and Modified day] July 21st ,2009

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